Ionic polymers as anti-infective agents

ABSTRACT

A method for treating a microbial infection in a mammal, such as a human, comprising treating the mammal with a therapeutically effective amount of a polymer comprising an amino group or an ammonium group attached to the polymer backbone via an aliphatic spacer group. The polymer can be a homopolymer or a copolymer. In one embodiment, the polymer is a copolymer comprising a monomer having a pendant ammonium group and a hydrophobic monomer.

RELATED APPLICATIONS

This application is a continuation application of U.S. Ser. No.09/286,693, filed Apr. 6, 1999, now abandoned which is a continuationapplication of U.S. Ser. No. 08/670,764, filed Jun. 24, 1996, now U.S.Pat. No. 6,034,129. The teachings of each of these referencedapplications are expressly incorporated herein by reference in theirentirety.

GOVERNMENT SUPPORT

The invention described herein was supported in whole or in part byAdvanced Technology Program Cooperative Agreement No. 70NANB5H1063 fromthe National Institute of Standards and Technology. The United StatesGovernment has certain rights in the invention.

BACKGROUND OF THE INVENTION

A number of short (ca. 50 amino acid residues or fewer) linear or cycliccytotoxic peptides have been isolated recently from a variety ofsources. These include mellitin, from bee venom, the magainins, fromfrog skin, and cecropins, from insects (Maloy, et al., Biopolymers(Peptide Science) 37: 105-122 (1995)). Although of widely varyingpeptide sequences and structures, these peptides all contain multiplelysine and arginine residues, and, at physiological pH, carry a netpositive charge. They also form amphipathic structures wherein oneportion of the structure is hydrophilic while the other portion ishydrophobic.

The peptides appear to act solely by direct lysis of the cell membrane(Maloy et al., supra (1995)). In the current model, cell lysis isinitiated by the electrostatic attraction of the positive charge on thepeptide to the negative phosphate head groups at the exterior surface ofthe membrane phospholipid bilayer. This interaction leads to insertionof the hydrophobic portion of the protein into the membrane, therebydisrupting the membrane structure. The lytic peptides are, in general,more active against prokaryotic cells, such as bacteria and fungi, thaneukaryotic cells. This has led to interest in these peptides aspotential agents for the treatment of infections in humans (Maloy etal., supra (1995); Arrowood et al., J Protozool. 38: 161S-163S (1991);Haynie et al., Antimicrob. Agents Chemotherapy 39: 301-307 (1995).

The natural cytotoxic peptides, however, suffer from severaldisadvantages with respect to their use as human therapeutic agents.First, it appears that these peptides have evolved to act at highconcentration at specific localized sites. Thus, when administered as adrug, the dosage necessary to attain an effective concentration at siteof infection can be prohibitively high. A second disadvantage is thedifficulty of isolating useful amounts of these peptides from thenatural sources, along with the high cost of synthesizing useful amountsof peptides in this size regime. Finally, these compounds, like otherpeptides, are degraded in the gastrointestinal tract, and, thus, cannotbe administered orally.

There is a need for anti-microbial agents which possess the broadactivity spectrum of the natural cytotoxic peptides, but are inexpensiveto produce, can be administered orally and have lower concentrationrequirements for therapeutic activity.

SUMMARY OF THE INVENTION

One aspect of the present invention is a method for treating a microbialinfection in a mammal, comprising administering to the mammal atherapeutically effective amount of a polymer having an amine orammonium group connected to the polymer backbone by an aliphatic spacergroup.

The polymer to be administered can be a homopolymer or a copolymer. Inone embodiment, the polymer further includes a monomer comprising ahydrophobic group, such as an aryl group or a normal or branchedC₃-C₁₈-alkyl group.

The polymer to be administered can, optionally, further include amonomer comprising a neutral hydrophilic group, such as a hydroxyl groupor an amide group.

Another aspect of the invention is a method for treating a microbialinfection in a mammal, such as a human, comprising administering to themammal a therapeutically effective amount of a polymer comprising apolymethylene backbone which is interrupted at one or more points by aquaternary ammonium group.

The present method has several advantages. For example, the polymersemployed are easily prepared using standard techniques of polymersynthesis and inexpensive starting materials. The polymers will not besubstantially degraded in the digestive tract and, therefore, can beadministered orally. Polymer compositions can also be readily varied, tooptimize properties such as solubility or water swellability andantimicrobial activity. Finally, the polymers to be administered includeamine or ammonium functional groups attached to the polymer backbone viaaliphatic spacer groups. The structural flexibility of such spacergroups minimizes backbone constraints on the interaction of the ammoniumgroups with anionic targets.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a method for preventing or treating amicrobial infection in a mammal, such as a human, by administering tothe mammal a therapeutically effective amount of a polymer comprising aplurality of amino or ammonium groups which are attached to the polymerbackbone via aliphatic spacer groups.

As used herein, a “therapeutically effective amount” is an amountsufficient to inhibit, partially or totally, a microbial infection or toreverse development of a microbial infection or prevent or reduce itsfurther progression. The term “polymer” refers to a macromoleculecomprising a plurality of repeat units or monomers. The term includeshomopolymers, which are formed from a singly type of monomer, andcopolymers, which are formed of two or more different monomers. A“terpolymer” is a copolymer formed from three different monomers. Theterm polymer, as used herein, is intended to exclude proteins, peptides,polypeptides and proteinaceous materials.

As used herein, the term “polymer backbone” or “backbone” refers to thatportion of the polymer which is a continuous chain, comprising the bondswhich are formed between monomers upon polymerization. The compositionof the polymer backbone can be described in terms of the identity of themonomers from which it is formed, without regard to the composition ofbranches, or side chains, off of the polymer backbone. Thus, apoly(acrylamide) polymer is said to have a poly(acrylamide) backbone,without regard to the substituents on the acrylamide nitrogen atom,which are components of the polymer side chains. Apoly(acrylarnide-co-styrene) copolymer, for example, is said to have amixed acrylamide/styrene backbone.

The term “polymer side chain” or “side chain” refers to the portion of amonomer which, following polymerization, forms a branch off of thepolymer backbone. In a homopolymer all of the polymer side chains areidentical. A copolymer can comprise two or more distinct side chains.When a side chain comprises an ionic unit, for example, the ionic unitdepends from, or is a substituent of, the polymer backbone, and isreferred to as a “pendant ionic unit”. The term “spacer group”, as usedherein, refers to a polyvalent molecular fragment which is a componentof a polymer side chain and connects a pendant moiety to the polymerbackbone. The term “aliphatic spacer group” refers to a spacer groupwhich does not include an aromatic unit, such as a phenylene unit.

The term “addition polymer”, as used herein, is a polymer formed by theaddition of monomers without the consequent release of a small molecule.A common type of addition polymer is formed by polymerizing olefinicmonomers, wherein monomers are joined by the formation of acarbon-carbon bonds between monomers, without the loss of any atomswhich compose the unreacted monomers.

The term “monomer”, as used herein, refers to both (a) a single moleculecomprising one or more polymerizable functional groups prior to orfollowing polymerization, and (b) a repeat unit of a polymer. Anunpolymerized monomer capable of addition polymerization, can, forexample, comprise an olefinic bond which is lost upon polymerization.

The quantity of a given polymer to be administered will be determined onan individual basis and will be determined, at least in part, byconsideration of the individual's size, the severity of symptoms to betreated and the result sought. The polymer can be administered alone orin a pharmaceutical composition comprising the polymer, an acceptablecarrier or diluent and, optionally, one or more additional drugs.

The polymers can be administered, for example, topically, orally,intranasally, or rectally. The form in which the agent is administered,for example, powder, tablet, capsule, solution, or emulsion, depends inpart on the route by which it is administered. The therapeuticallyeffective amount can be administered in a series of doses separated byappropriate time intervals, such as hours.

Microbial infections which can be treated or prevented by the method ofthe present invention include bacterial infections, such as infection byStreptococcus, including Streptococcus mutans, Streptococcus salivarius,and Streptococcus sanguis, Salmonella, Campylobacter, includingCampylobacter sputum, Antinomyces, including Actinomyces naeslundii andActinomyces viscosus, Escherichia coli, Clostridium difficile,Staphylococcus, including S. aureus, Shigella, Pseudomonas, including P.aeruginosa, Eikenella corrodens, Actinobacillus actinomycetemcomitans,Bacteroides gingivalis, Capnocytophaga, including Capnocytophagagingivalis, Wolinell recta, Bacteriodes intermedius, Mycoplasma,including Mycoplasma salivarium, Treponema, including Treponemadenticola, Peptostreptococcus micros, Bacteriodes forsythus,Fusobacteria, including Fusobacterium nucleatum, Selenomonas sputigena,Bacteriodes fragilis, Enterobacter cloacae and Pneumocystis. Alsoincluded are protozoal infections, such as infection by Cryptosporidiumparvum and Giardia lamblia; ameobic infections, such as infection byEntameoba histolytica or Acanthameoba; fungal infections, such asinfections by Candida albicans and Aspergillus fumigatus, and arasiticinfections, such as infections by A. castellani and Trichinellaspiralis. The method is useful for treating infections of various organsof the body, but is particularly useful for infections of the skin andgastrointestinal tract.

Polymers which are particularly suitable for the present method includepolymers which can possess key characteristics of naturally occurringcytotoxic peptides, in particular, the ability to form amphipathicstructures. The term “amphipathic”, as used herein, describes athree-dimensional structure having discrete hydrophobic and hydrophilicregions. Thus, one portion of the structure interacts favorably withaqueous and other polar media, while another portion of the structureinteracts favorably with non-polar media. An amphipathic polymer resultsfrom the presence of both hydrophilic and hydrophobic structuralelements along the polymer backbone.

In one embodiment, the polymer to be administered polymer comprises amonomer of Formula I,

wherein X is a covalent bond, a carbonyl group or a CH₂ group, Y is anoxygen atom, an NH group or a CH₂ group, Z is an spacer group, R is ahydrogen atom or a methyl or ethyl group, R₁, R₂ and R₃ are each,independently, a hydrogen atom, a normal or branched, substituted orunsubstituted C₁-C₁₈-alkyl group, an aryl group or an arylalkyl group.Suitable alkyl substituents include halogen atoms, such as fluorine orchlorine atoms.

In the case in which at least one of R₁-R₃ is a hydrogen atom, themonomer can also exist in the free base, or amino form, that is, as theneutral conjugate base of the ammonium cation. The polymer comprisingsuch a monomer can be administered in the protonated, cationic form,such as a salt of a pharmaceutically acceptable acid, or in the freebase form. Suitable acids include hydrochloric acid, hydrobromic acid,citric acid, lactic acid, tartaric acid, phosphoric acid,methanesulfonic acid, acetic acid, formic acid, maleic acid, fumaricacid, malic acid, succinic acid, malonic acid, sulfuric acid, L-glutamicacid, L-aspartic acid, pyruvic acid, mucic acid, benzoic acid,glucoronic acid, oxalic acid, ascorbic acid, and acetylglycine. Ineither case, at physiological pH following administration, a pluralityof amino groups will be protonated to become ammonium groups, and thepolymer will carry an overall positive charge.

The spacer group is a component of the polymer side chain and connectsthe amino or ammonium group to the polymer backbone. The amino orammonium group is, thus, a pendant group. The spacer group can be anormal or branched, saturated or unsaturated, substituted orunsubstituted alkylene group, such as a polymethylene group —(CH₂)_(n)—,wherein n is an integer from about 2 to about 15. Suitable examplesinclude the propylene, hexylene and octylene groups. The alkylene groupcan also, optionally, be interrupted at one or more points by aheteroatom, such as an oxygen, nitrogen (e.g, NH) or sulfur atom.Examples include the oxaalkylene groups —(CH₂)₂O[(CH₂)₂O]_(n)(CH₂)₂-,wherein n is an integer ranging from 0 to about 3.

Examples of monomers of Formula I having quaternary ammonium groupsinclude 2-trimethylammonium-ethylmethacrylate,2-trimethylammoniumethylacrylate,N-(3-trimethylammoniumpropyl)methacrylamide,N-(6-trimethylammoniumhexyl)acrylamide,N-(3-trimethylammoniumpropyl)acrylamide, andN-(4-trimethylammoniumbutyl)allylamine, each of which also includes acounter anion. Examples monomers of Formula I having an amino groupinclude allylamine and N-(3-dimethylaminopropyl)acrylamide.

Polymers to be administered which have quaternary ammonium groups orprotonated amino groups will further comprise a pharmaceuticallyacceptable counter anion, such as anions which are conjugate bases ofthe pharmaceutically acceptable acids discussed above, for example,chloride, bromide, acetate, formate, citrate, ascorbate, sulfate orphosphate. The number of counter anions associated with the polymerprior to administration is the number necessary to balance theelectrical charge on the polymer.

The polymer can also be a copolymer further comprising a hydrophobicmonomer. The hydrophobic monomer can comprise a side chain bearing ahydrophobic group, such as a straight chain or branched, substituted orunsubstituted C₃-C₁₈-alkyl group or a substituted or unsubstituted arylgroup. Examples of suitable hydrophobic monomers include styrene,N-isopropylacrylamide, N-t-butylacrylamide, N-n-butylacrylamide,heptafluorobutylacrylate, N-n-decylallylamine, N-n-decylacrylamide,pentafluorostyrene, n-butylacrylate, t-butylacrylate, n-decylacrylate,N-t-butylmethacrylamide, n-decylmethacrylate, and n-butylmethacrylate.

Examples of copolymers comprising a monomer of Formula I and ahydrophobic monomer includepoly(N-(3-dimethylaminopropyl)acrylamide-co-N-(n-butyl)acrylamide) orsalts thereof with pharmaceutically acceptable acids. Other examples ofsuitable copolymers includepoly(2-trimethylammoniumethylmethacrylate-co-styrene) chloride,poly(2-trimethylammoniumethyhnethacrylate-co-N-isopropylacrylamide)chloride,poly(2-trimethyl-ammoniumethylmethacrylate-co-heptafluorobutylacryl)chloride, poly(3-trimethylammoniumpropylmethacrylate-co-styrene)chloride,poly(3-trimethylammonium-propylmethacrylate-co-N-t-butylacrylamide)chloride,poly(3-trimethylaminoniumpropylmethacrylate-co-N-n-butylacrylamide)chloride, andpoly(N-(3-trimethylammoniumpropyl)allylamine-co-N-n-decylallylamine).Each of these ionic copolymers can also be employed with counter ionsother than chloride, for example, a conjugate base of a pharmaceuticallyacceptable acid.

In a further embodiment, the polymer to be administered comprises amonomer of Formula I, a hydrophobic monomer and a neutral hydrophilicmonomer, such as acrylamide, methacrylamide,N-(2-hydroxyethyl)acrylamide or 2-hydroxyethylmethacrylate. Examples ofpolymers of this type include terpolymers ofN-(3-trimethylammonium-propyl)methacrylamide/N-isopropylacrylamide/2-hydroxyethyl-methacrylate,N-(3-trimethylammonium-propyl)methacrylamide/N-n-decylacrylamide/2-hydroxyethylmethacrylate,N-(3-trimethylammoniumpropyl)methacrylamide/N-t-butylmethacrylamide/methacrylamide,N-(3-trimethylammonium-propyl)methacrylamide/n-decylacrylate/methacrylamide,2-trimethylammoniumethylmethacrylate/n-butyl-acrylate/acrylamide,2-trimethylammonium-ethylmethacrylate/t-butylacrylate/acrylamide,2-trimethylammoniumethylmethacrylate/n-decyl-acrylate/acrylamide,2-trimethylammonium-ethylmethacrylate/n-decylmethacrylate/methacrylamide,2-trimethylammoniumethylmethacrylate/N-t-butyl-methacrylamide/methacrylamideand2-trimethylammoniumethylmethacrylate/N-n-butyl-methacrylamide/methacrylamide.

The polymer to be administered can be an addition polymer having apolymer backbone such as a polyacrylate, polyacrylamidepoly(allylalcohol), poly(vinylalcohol), poly(vinylamine),poly(allylamine), or polyalkyleneimine backbone. The polymer can have auniform backbone if it is composed of monomers derived from a commonpolymerizable unit, such as acrylamide. If the polymer is a copolymer,it can also comprise a mixed backbone, for example, the monomer ofFormula I can be an acrylamide derivative, while the hydrophobic monomercan be a styrene derivative. The polymers disclosed herein includeexamples of both uniform and mixed backbones.

The polymers of use in the present method also include condensationpolymers, wherein polymerization of monomers is accompanied by therelease of a small molecule, such as a water molecule. Such polymersinclude, for example, polyesters and polyurethanes.

The polymers of use in the present method are preferably substantiallynonbiodegradable and nonabsorbale. That is, the polymers do notsubstantially break down under physiological conditions into fragmentswhich are absorbable by body tissues. The polymers preferably have anonhydrolyzable backbone, which is substantially inert under conditionsencountered in the target reion of the body, such as thegastrointestinal tract.

The composition of the copolymers to be administered can varysubstantially. The copolymer can comprise from about 95 mole percent toabout 5 mole percent, preferably from about 20 mole percent to about 80mole percent, of a monomer of Formula I. The copolymer can also comprisefrom about 95 mole percent to about 5 mole percent, preferably fromabout 20 mole percent to about 80 mole percent, of a hydrophobicmonomer.

Other examples of polymers which are of use in the present method aredisclosed in U.S. patent application Ser. Nos. 08/482,969, 08/258,477,08/258,431, 08/469,659 and 08/471,769, the contents of each of which areincorporated herein by reference.

The polymer to be administered will, preferably, be of a molecularweight which is suitable for the intended mode of administration andallows the polymer to reach and remain within the targeted region of thebody for a period of time sufficient to interact with the infectingorganism. For example, a method for treating an intestinal infectionshould utilize a polymer of sufficiently high molecular weight to resistabsorption, partially or completely, from the gastrointestinal tractinto other parts of the body. The polymers can have molecular weightsranging from about 500 Daltons to about 500,000 Daltons, preferably fromabout 2,000 Daltons to about 150,000 Daltons.

The polymers which are useful in the present method can be prepared byknown methods. A first method includes the direct polymerization of amonomer, such as trimethylammoniumethylacrylate chloride, or a set oftwo or more monomers, such as trimethylammoniumethyl-acrylate chloride,N-n-butylacrylamide and acrylamide. This can be accomplished viastandard methods of free radical, cationic or anionic polymerizationwhich are well known in the art. Due to reactivity differences betweentwo monomers, the composition of a copolymer produced in this way candiffer from the composition of the starting mixture. This reactivitydifference can also result in a non-random distribution of monomersalong the polymer chain.

A second method proceeds via the intermediacy of an activated polymercomprising labile side chains which are readily substituted by a desiredside chain. An example of a suitable activated polymer is thesuccinimide ester of polyacrylic acid, poly(N-acryloyloxysuccinimide)(also referred to hereinafter as “pNAS”), which reacts with nucleophilessuch as a primary amine to form a N-substituted polyacrylamide. Anothersuitable activated polymer is poly(para-nitrophenylacrylate), whichreact with amine nucleophiles in a similar fashion.

Polymers suitable for use in the present method can also be prepared byaddition of a side chain to a preformed polymer. For example,poly(allylamine) can be alkylated at the amino nitrogen by one or morealkylating agents. For example, one fraction of amino groups can bealkylated using a normal or branched C₃-C₁₈-alkyl halide, such asn-decyl bromide, while another fraction can be alkylate by a quaternaryammonium-containing alkyl halide, such as1-trimethylammonium-4-bromombutane.

A copolymer having a polyacrylamide backbone comprising amide nitrogensbearing two different substituents can be prepared by treating p(NAS)with less than one equivalent (relative to N-acryloyloxysuccinimidemonomer) of a first primary amine, producing a poly(N-substitutedacrylamide-co-N-acryoyloxysuccinimide) copolymer. RemainingN-acryoyloxysuccinimide monomer can then be reacted with, for example,an excess of a second primary amine to produce a polyacrylamidecopolymer having two different N-substituents. A variety of copolymercompositions can, thus, be obtained by treating the activated polymerwith different proportions of two or more amines.

An additional aspect of the present invention is a method for treating amicrobial infection in a mammal, such as a human, comprisingadministering to the mammal a therapeutically effective amount of apolymer having an amino group or an ammonium group within the polymerbackbone. The polymer can have, for example, a polymethylene backbonewhich is interrupted by one or more amino or ammonium groups. An exampleof a polymer of this type ispoly(decamethylenedimethylammonium-co-ethylenedimethylammonium) bromide,which is synthesized via the reaction ofN,N,N′,N′-tetramethylethylenediamine and 1,10-dibromodecane. The polymercan also be administered in association with anions other than bromide,such as chloride or acetate anions. Other examples includepoly(alkyleneimines), for example, poly(ethyleneimine). Such polymerscan comprise secondary or tertiary amino groups, salts of such groupswith pharmaceutically acceptable acids, and/or quaternary ammoniumgroups.

As discussed below in Example 35, several polymers described herein weretested for in vitro activity against Cryptosporidium parvum infectivityin mammalian ell culture. Of these, poly(TMAEMC-co-styrene), describedin Example 7, was most active, exhibiting greater than 90% inhibition ofC. parvum infectivity relative to the control when applied as a 0.1mg/mL, solution in dimethylsulfoxide. The remaining polymers tested alsoshowed significant anti-Cryptosporidium activity.

The invention will now be further and specifically described by thefollowing examples.

EXAMPLES

The following abbreviations are used throughout the examples to denotethe following monomers: MAPTAC,N-(3-trimethylammoniumpropyl)methacrylamide chloride; TMAEMC,2-trimethylammoniumethylmethacrylate chloride; HEMA,2-hydroxyethylmethacrylate; TMAEAC, 2-trimethylammoniumethylacrylatechloride.

The copolymers and terpolymers of the following examples are givennominal compositions which correspond to the molar ratios of startingmonomers in the copolymerization mixture.

Example 1 Synthesis of poly(N-acryloyloxysuccinimide) (PNAS)

A solution of N-acryloyloxysuccinimide (25.0 g, 148 mmole) in 100 mL dryDMF was degassed by nitrogen purging and simultaneously heated to 60° C.To the reaction mixture was added azobisisobutyronitrile (AIBN) (120 mg,0.005 equivalents with respect to monomer). The reaction was allowed toproceed for 24 hours at 60° C. The polymer solution was cooled to roomtemperature and poured into rapidly stirred THF. The resulting whiteprecipitate was filtered, washed with THF and dried in vacuo.

Example 2 Synthesis ofpoly(N-(3-dimethylamino-propyl)acrylamide-co-N-n-butylacrylamide)

To a solution of 3.0 g (17.75 mmole) pNAS in 20 mL dry DMF was added 0.6g (3.55 mmole) n-butylamine. The resulting solution was stirred at roomtemperature for 14 hours, and then heated at 60° C. for 4 hours. Afterthe solution was cooled to room temperature, 9.05 g (89 mmole)3-dimethylaminopropylamine was added, and the resulting solution wasstirred at room temperature for 2 hours, then heated to 60° C. for 20hours. After cooling to room temperature, the solution was diluted with25 mL water, and dialyzed against water for 24 hours. The solution wasthen lyophilized to affordpoly(N-(3-dimethylaminopropyl-acrylamide)-co-N-n-butylacrylamide) as atacky white solid.

Example 3 Synthesis ofpoly(N-(3-trimethylammoniumpropyl)acrylamide-co-N-n-butylacrylamide)iodide

To a suspension ofpoly(3-dimethylaminopropyl-acrylamide-co-N-n-butylacrylamide in methanolwas added 0.5 g methyl iodide. The resulting mixture was stirred for 3hours, and gradually became homogeneous. After stirring for another 12hours, the solvent was removed under reduced pressure and the polymerwas washed with dry hexane.

Example 4 Synthesis ofpoly(N-(2-hydroxyethyl)acrylamide-co-N-(6-trimethylammoniumhexyl)acrylamide)bromide

To a solution of 2.48 g (15 mmole) pNAS in 5 mL DMF was added 1.00 g (3mmole) 1 -trimethylammonium-6-hexanamine bromide. The solution wasstirred at room temperature for 4 hours and then heated at 60° C. for 20hours. The solution was cooled to room temperature, and then 8.95 g (150mmole) 2-ethanolamine was added. The resulting mixture was heated to 80°C. for 20 hours, cooled to room temperature and diluted with 10 mLwater. The solution was dialyzed against water for 24 hours, thenlyophilized, yielding the polymer as a brittle white solid.

Example 5 Synthesis of poly(TMAEAC)

A solution of 48.25 g (0.25 mol) 2-trimethylammoniumethylacrylatechloride in 400 mL isopropanol was degassed by nitrogen purging andheated to 35° C. To this stirred solution was added a solution of 0.8 gpotassium persulfate in 10 mL distilled water. A slight exotherm wasobserved. The solution was stirred at 35° C. for 6 hours, then cooled toroom temperature. The solution was added to hexanes and the resultingprecipitate was isolated by filtration.

Example 6 Synthesis ofpoly(decamethylenedimethylammonium-co-ethylenedimethylammonium) bromide

N,N,N′N′-tetramethylethylenediamine (10.0 g, Aldrich),1,10-dibromodecane (25.8 g, Aldrich) and methanol (100 mL) were placedinto a three-neck 250 mL round bottom flask. The mixture was heated withgentle stirring to 65° C. for 6 days, at which point methanol (40 mL)was added, and the mixture was refluxed for an additional 2 days. Themixture was then dripped into acetone, forming a solid that wascollected by filtration, rinsed with acetone, and dried in a vacuum ovento yield 30.9 g of product.

Example 7 Synthesis of poly(TMAEMC-co-styrene) 75/25

A 500 mL round bottomed flask was charged withtrimethylammoniumethylmethacrylate chloride (26.0 g of a 70 wt % aqueoussolution, 18.2 g), styrene (6.0 g) and isopropanol (150 mL). Thesolution was degassed by the addition of a rapid stream of nitrogen for10 minutes, followed by the addition of AIBN (0.5 g). The solution wasdegassed for a further thirty minutes and, while continuing the additionof nitrogen, the solution was heated to 70° C., and the temperaturemaintained for 17 h. The polymer began to precipitate within 2 h, and bythe completion of the reaction a sticky white precipitate had formed.The reaction mixture was cooled, the isopropanol was decanted from thepolymer, and the polymer was dissolved in methanol. Dropwise addition ofthe methanol solution to ethyl acetate (1200 mL) caused the polymer toprecipitate as a fine white powder which was recovered by filtration.

Example 8 Synthesis of poly(TMAEMC-co-N-isopropylacrylamide) (67/33)

A 500 mL round bottomed flask was charged withtrimethylammoniumethylmethacrylate chloride (14.5 g of a 70 wt % aqueoussolution, 10.0 g), N-isopropylacrylamide (5.0 g) and isopropanol (150mL). The solution was degassed by the addition of a rapid stream ofnitrogen for 10 minutes, followed by the addition of AIBN (0.5 g). Thesolution was degassed for a further 60 minutes. The reaction mixture washeated to 70° C., and the temperature maintained for 16 h. The polymerpartially precipitated over the course of the reaction. Upon cooling,the propanol was decanted from the polymer, and the polymer wasdissolved in methanol. Precipitation of the methanol solution dropwiseinto ethyl acetate (1200 mL) caused the polymer to be deposited as whitecurds which were recovered by filtration, washed with ethyl acetate, anddried in vacuo.

Additional TMAEMC/N-isopropylacrylamide copolymers were prepared by asimilar method with the starting monomers in the following ratios:TMAEMC/N-isopropylacrylamide=40/60, 25/75 and 15/85.

Example 9 Synthesis of poly(MAPTAC-co-styrene) 75/25

To isopropanol (150 mL) was added a solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride in water (50 wt %solution, 24.0 g, 12.0 g of monomer). To this solution was added styrene(6.0 g), followed by the addition of AIBN (0.5 g). The homogeneoussolution was degassed by bubbling a stream of nitrogen through it for 30minutes. The solution was heated to 70° C. for 15 h. The polymerpartially precipitated as the reaction proceeded. The solution wascooled, the isopropanol was decanted off, the white solid was washedwith propanol (50 mL). The propanol was decanted a second time, and thesolid was dissolved in methanol (150 mL). The clear solution was addeddropwise to ethyl acetate, causing the polymer to be precipitated as awhite powder. The polymer was recovered by filtration, washed with 50 mLof ethylacetate and air dried.

An additional MAPTAC/styrene copolymer was prepared by a similar methodemploying a 50/50 mixture of starting monomers.

Example 10 Synthesis of poly(TMAEMC-co-heptafluorobutylacrylate) 75/25

A 500 mL round bottomed flask was charged with2-trimethylammoniumethylmethacrylate chloride (26.0 g of a 70wt %aqueous solution, 18.2 g), heptafluorobutylacrylate (6.0 g) andisopropanol (150 mL). The solution was degassed by the addition of arapid stream of nitrogen for 10 minutes, followed by the addition ofAIBN (0.5 g)..The solution was degassed for a further thirty minutesand, continuing the addition of nitrogen, the solution was heated to 70°C. The temperature was maintained for 17 h. The polymer began toprecipitate within 1 h, and by the completion of the reaction a stickywhite precipitate had formed. The reaction mixture was cooled, thepropanol was decanted from the polymer, and the polymer was dissolved inmethanol (100 mL). Precipitation of the methanol solution dropwise intoethyl acetate (1200 mL) caused the polymer to be deposited as a whitesolid which was recovered by filtration.

Example 11 Synthesis of poly(MAPTAC-co-N-t-butylacrylamide) 75/25

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 36.4 g of a 50% aqueous solutionof N-(3-trimethylammonium-propyl)methacrylamide chloride and 6 g ofN-t-butyl-acrylamide followed by 150 mL of isopropanol. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for 15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The resulting reaction mixture consisted of two phases. The turbidliquid phase was decanted from the bulk of the reaction which was awhite sticky solid phase. The liquid was precipitated into 1200 mL ofethyl acetate and filtered by vacuum filtration through a Buchnerfunnel. The white hygroscopic precipitate was dried in vacuo. The solidphase was dissolved in methanol and precipitated into 1200 mL of ethylacetate and filtered by vacuum filtration to yield a white powder whichwas stored under vacuum.

Additional MAPTAC/N-t-butylacrylamide copolymers were prepared by asimilar method beginning with the starting monomers in the followingratios:N-(3-trimethylammoniumpropyl)methacrylamide/N-t-butyl-acrylamide=60/40,50/50, 40/60, and 25/75

Example 12 Synthesis ofpoly(N-decylallylamine-co-N-(4-trimethylammoniumbutyl)allylamine)

To a solution of poly(allylamine).HCl (20.15 g of a 50 wt % aqueoussolution) was added sodium hydroxide (5.64 g ) as a solid. The solutionwas stirred for 40 minutes, filtered and the filter cake was washed withmethanol (15 mL). The solution was further diluted with methanol (25 mL)and to the solution was added 1-bromodecane (7.73 g, 35 mmol) and(1-trimethylamino-4-bromobutane) chloride (9.13 g, 35 mmol). A solutionwas prepared of sodium hydroxide (2.8 g, 70 mmol) in water (5 mL). Thissolution was added to the reaction mixture in four portions at thirtyminute intervals. The solution was then stirred at room temperature for24 h, followed by dialysis against deionized water and freeze-dried. Atotal of 23.2 g of a glassy, hygroscopic solid was recovered.

Example 13 Synthesis of poly(TMAEMC-co-N-t-butylacrylamide) 57/43

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 18.20 g of a 70% aqueous solutionof 2-trimethylammonium- ethylmethacrylic chloride and 9.7 g ofN-t-butylacrylamide followed by 150 mL of isopropanol. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for ˜15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The resulting reaction mixture consisted of two easily separable phases.The liquid phase was decanted from the bulk of the reaction which was awhite solid. The liquid was precipitated into 1200 mL of ethyl acetateand filtered by vacuum filtration through a Buchner funnel. The whiteprecipitate was dried in vacuo and weighed: fraction A, 1 0.1 g (45.1 %yield based on 22.4 g monomers added). The solid phase was dissolved inmethanol and precipitated into 600 mL of ethyl acetate and filtered byvacuum filtration to yield fraction B, 5.81 g of a white powder (25.9%yield) which was dried under vacuum.

TMAEMC/N-t-Butylacrylamide copolymers were also prepared by a similarmethod with the starting monomers in the following ratios:TMAEMC/N-t-Butylacrylamide=63/37, 50/50, 40/60, 25/75, 15/85 and 5/95.

Example 14 Synthesis of poly(MAPTAC-co-N-n-decylacrylamide) 75/25

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 36.4 g of a 50% aqueous solutionof N-(3-trimethylammoniumpropyl)methacrylamide chloride and 6 g ofN-n-decylacrylamide followed by 150 mL of isopropanol. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for ˜15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of two easily separable phases. Theclear, yellow liquid phase was precipitated into 1200 mL of ethylacetate. The precipitate was isolated by filtration and dried undervacuum to yield 2.14 g of a yellow powder, fraction A (8.84% yield).Methanol was added to the creamy yellow reaction precipitate and theresulting turbid yellow solution was precipitated into 1200 mL of ethylacetate. The white precipitate was isolated by filtration and driedunder vacuum to yield fraction B, 17.22 g, as a slightly yellow powder(71.2% yield).

Additional MAPTAC/N-n-decylacrylamide copolymers were prepared by asimilar method with the starting monomers in the following ratios:MAPTAC/N-n-decylacrylamide=60/40, 50/50, and 40/60.

Example 15 Synthesis of poly(TMAEMC-co-pentafluorostyrene) 75/25

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 26.0 g of a 70% aqueous solutionof 2-trimethylammonium-ethylmethacrylate chloride and 6 g ofpentafluorostyrene followed by 150 mL of isopropanol. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for ˜15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of two phases. The turbid solution wasdiscarded. The bulk of the reaction, consisting of a white solid mass atthe bottom of the flask, was dissolved in methanol. The resulting clearsolution was precipitated into 1200 mL of ethyl acetate. The whiteprecipitate was isolated by vacuum filtration to yield 20.39 g of a finewhite powder (84.3% yield).

Additional TMAEMC/pentafluorostyrene copolymers were prepared by asimilar method with the starting monomers in the following ratios:TMAEMC/pentafluorostyrene =60/40 and 50/50.

Example 16 Synthesis of poly(MAPTAC-co-pentafluorostyrene) 75/25

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 36.3 g of a 50% aqueous solutionof N-(3-trimethylammoniumpropyl)methacrylamide chloride and 6 g ofpentafluorostyrene followed by 150 mL of isopropanol. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for ˜15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of a turbid solution with a whiteprecipitate. The supernatent was disgarded. The white reactionprecipitate was dissolved in methanol and the resulting clear solutionwas precipitated into 1200 mL of ethyl acetate. The white precipitatewas isolated by filtration and dried under vacuum to yield 12.81 g of afine white powder (52.9% yield).

Additional MAPTAC/pentafluorostyrene copolymers were prepared by asimilar method with the starting monomers in the following ratios:MAPTAC/pentafluorostyrene=60/40 and 50/50.

Example 17 Synthesis of MAPTAC/N-t-Butylacrylamide/HEMA Terpolymer33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.1 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 8 g ofN-t-butylacrylamide, and 8 g of 2-hydroxyethylmethacrylate. The solutionwas purged with nitrogen for 1 hour and 0.5 g of AIBN was added. Themixture was purged for ˜15 min until all of the AIBN dissolved. Thesolution was heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of a turbid solution with a white latexin the bottom of the flask. The solution was precipitated into 1200 mLof ethyl acetate. The white precipitate was isolated by filtration toyield a sticky white powder which was dried under vacuum to yield 10.43g of a lumpy white solid (fraction A) (43.1% yield). The white reactionprecipitate was dissolved in methanol and precipitated into 1200 mL ofethyl acetate. The precipitate was isolated by filtration and driedunder vacuum to yield 8.89 g of a fine white powder (fraction B) (36.7%yield).

Additional MAPTAC/N-t-butylacrylamide/HEMA terpolymers were prepared bya similar method beginning with the following ratios of the startingmonomers: MAPTAC/N-t-Butylacrylamide/HEMA=28/43/28, 23/53/23, and18/63/18.

Example 18 Synthesis of MAPTAC/N-Isopropylacrylamide/HEMA Terpolymer18/63/18

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by8.9 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 15.3 g ofN-iso-propylacrylamide, and 4.4 g of 2-hydroxyethylmethacrylate. Thesolution was purged with nitrogen for 1 hour and 0.5 g of AIBN wasadded. The mixture was purged for ˜15 min until all of the AIBNdissolved. The solution was heated to 75° C. under nitrogen for 16hours.

The clear slightly pink reaction solution was precipitated into 1200 mLof ethyl acetate. The precipitate was isolated by filtration to yield asticky white solid which was dried under vacuum to yield 14.42 g of ahard clear/white granular solid (59.6% yield).

Example 19 Synthesis of MAPTAC/N-Decylacrylamide/HEMA Terpolymer33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.1 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 8 g ofN-decylacrylamide, and 8 g of 2-hydroxyethylmethacrylate. The solutionwas purged with nitrogen for 1 hour and 0.5 g of AIBN was added. Themixture was purged for ˜15 min until all of the AIBN dissolved. Thesolution was heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of two phases. The clear yellow solutionwas precipitated into 1200 mL of ethyl acetate. The precipitate wasisolated by filtration. The sticky yellow precipitate was dried undervacuum and the resulting brittle clear yellow foam was crushed to yield4.98 g of a fine yellow granular powder (fraction A) (20.6% yield). Thewhite reaction latex was dissolved in methanol and precipitated into1200 mL of ethyl acetate. The precipitate was isolated by filtration anddried under vacuum to yield 10.24 g of a slightly yellow granular solid(fraction B) (42.3% yield).

Additional MAPTAC/N-Decylacrylamide/HEMA terpolymers were prepared by asimilar method beginning with the following ratios of starting monomers:MAPTAC/N- Decylacrylamide/HEMA=28/43/28, 23/53/23, and 18/63/18.

Example 20 Synthesis of TMAEAC/n-Butylacrylate/Acrylamide Terpolymer10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 7.26 g of n-butylacrylate, and 14.52 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The resulting white reaction mixture was filtered by vacuum filtrationthrough a Buchner funnel to yield a white powder. The powder was washedwith isopropanol and dried under vacuum to yield 21.57 g of a fine whitepowder (89.1% yield based on 24.2 g of monomers).

Additional TMAEAC/n-butylacrylate/acrylamide terpolymers were preparedby a similar method beginning with the following ratios of startingmonomers: TMAEMC/n-butylacrylate/acrylamide =20/20/60 and 30/10/60.

Example 21 Synthesis of TMAEAC/t-Butylacrylate/Acrylamide Terpolymer10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 7.26 g of t-butylacrylate, and 14.52 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The resulting white reaction mixture was filtered by vacuum filtrationthrough a Buchner funnel to yield a white powder. The powder was washedwith isopropanol and dried under vacuum to yield 21.13 g of a whitepowder (87.3% yield).

Additional TMAEAC/t-butylacrylate /acrylamide terpolymers were preparedby a similar method beginning with the following ratios of startingmonomers: TMAEAC/t-butylacrylate/acrylamide=20/20/60 and 30/10/60.

Example 22 Synthesis of TMAEAC/n-Decylacrylate/Acrylamide Terpolymer10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 7.26 g of n-decylacrylate, and 14.52 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The resulting white reaction mixture was filtered by vacuum filtrationthrough a Buchner funnel to yield a white powder. The powder was washedwith isopropanol and dried under vacuum to yield 21.52 g of a fine whitepowder (89% yield).

Additional TMAEAC/n-decylacrylate /acrylamide terpolymers were preparedby a similar method beginning with the following ratios of startingmonomers: TMAEAC/n-decylacrylate/acrylamide=20/20/60, and 30/10/60.

Example 23 Synthesis of MAPTAC/N-t-Butylmethacrylamide/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 7.26 g ofN-t-butylmethacrylamide, and 14.52 g of methacrylamide. The solution waspurged with nitrogen for 1 hour and 0.5 g of AIBN was added. The mixturewas purged for ˜15 min until all of the AIBN dissolved. The solution washeated to 75° C. under nitrogen for 16 hours.

The white reaction mixture was too difficult to filter by vacuumfiltration so centrifugation techniques were employed instead. Thereaction mixture was poured into 50 mL centrifuge tubes and centrifuged.The supernatant was discarded. Isopropanol was added to the polymer andthe mixture was stirred and centrifuged. The supernatant was discardedand the white solids were combined and dried under vacuum to yield 14.99g of a slightly buff powder (61.9% yield).

Additional MAPTAC/N-t-butylmethacrylamide/ methacrylamide terpolymerswere prepared by a similar method beginning with the following ratios ofstarting monomers: MAPTAC/N-t-butylmethacrylamide/methacrylamide20/20/60, 33/33/33 and 30/10/60.

Example 24 Synthesis of MAPTAC/n-Decylmethacrylate/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 7.26 g ofn-decylmethacrylate, and 14.52 g of methacrylamide. The solution waspurged with nitrogen for 1 hour and 0.5 g of AIBN was added. The mixturewas purged for ˜15 min until all of the AIBN dissolved. The solution washeated to 75° C. under nitrogen for 16 hours.

The isopropanol was decanted leaving a white chunky powder. Isopropanolwas added and the mixture was poured into 50 mL centrifuge tubes andcentrifuged. The supernatant was discarded. Isopropanol was added to thepolymer and the mixture was stirred and centrifuged. The supernatant wasdiscarded and the white solids were combined and dried under vacuum toyield 18.50 g of a granular white solid (76.4% yield).

Additional MAPTAC/N-decylmethacrylamide/methacrylamide terpolymers wereprepared by a similar method beginning with the following ratios ofstarting monomers: MAPTAC/N-decylmethacrylamide/methacrylamide=20/20/60,33/33/33 and 30/10/60.

Example 25 Synthesis of TMAEMC/n-Decylmethacrylate/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by3.46 g of a 70% aqueous solution of 2-trimethylammoniumethylmethacrylatechloride, 7.26 g of n-decylmethacrylate, and 14.52 g of methacrylamide.The solution was purged with nitrogen for 1 hour and 0.5 g AIBN wasadded. The mixture was purged for ˜15 minutes until all of the AIBNdissolved. The solution was heated to 75° C. under nitrogen for 16hours.

The white reaction mixture was poured into 50 mL centrifuge tubes andcentrifuged. The supernatant was discarded. Isopropanol was added to thepolymer and the mixture was stirred and centrifuged. The supernatant wasdiscarded and the white solids were combined and dried under vacuum toyield 10.29 g of a hard white solid (42.5% yield).

Additional TMAEMC/N-n-decylmethacrylamide/ methacrylamide terpolymerswere prepared by a similar method beginning with the following ratios ofstarting monomers:TMAEMC/N-n-decylmethacrylamide/methacrylamide=20/20/60, 33/33/33 and30/10/60.

Example 26 Synthesis of TMAEMC/N-t-Butylmethacrylamide/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by3.46 g of a 70% aqueous solution of 2-trimethylammoniumethylmethacrylatechloride, 7.26 g of N-t-butylmethacrylamide, and 14.52 g ofmethacrylamide. The solution was purged with nitrogen for 1 hour and 0.5g AIBN was added. The mixture was purged for ˜15 minutes until all ofthe AIBN dissolved. The solution was heated to 75° C. under nitrogen for16 hours.

The white reaction mixture was poured into 50 mL centrifuge tubes andcentrifuged. The supernatant was discarded. Isopropanol was added to thepolymer and the mixture was stirred and centrifuged. The supernatant wasdiscarded and the white solids were combined and dried under vacuum toyield 18.35 g of a fine white powder (75.8% yield).

Additional TMAEMC/N-t-butylmethacrylamide/ methacrylamide terpolymerswere prepared by a similar method beginning with the following ratios ofstarting monomers: TMAEMC/N-t-butylmethacrylamide/methacrylamide20/20/60, 33/33/33 and 30/10/60.

Example 27 Synthesis of TMAEMC/n-Butylmethacrylate/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by3.46 g of a 70% aqueous solution of 2-trimethylammoniumethylmethacrylatechloride, 7.26 g of n-butylmethacrylate, and 14.52 g of methacrylamide.The solution was purged with nitrogen for 1 hour and 0.5 g AIBN wasadded. The mixture was purged for ˜15 minutes until all of the AIBNdissolved. The solution was heated to 75° C. under nitrogen for 16hours.

The white reaction mixture was poured into 50 mL centrifuge tubes andcentrifuged. The supernatant was discarded. Isopropanol was added to thepolymer and the mixture was stirred and centrifuged. The supernatant wasdiscarded and the white solids were combined and dried under vacuum toyield 20.99 g of a clumpy white powder (86.7% yield).

Additional TMAEMC/N-n-butylmethacrylamide/ methacrylamide terpolymerswere prepared by a similar method beginning with the following ratios ofstarting monomers:TMAEMC/N-n-butylmethacrylamide/methacrylamide=20/20/60 and 30/10/60.

Example 28 Synthesis of MAPTAC/n-Butylmethacrylate/MethacrylamideTerpolymer 10/30/60

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by4.84 g of a 50% aqueous solution ofN-(3-trimethylammoniumpropyl)methacrylamide chloride, 7.26 g ofn-butylmethacrylate, and 14.52 g of methacrylamide. The solution waspurged with nitrogen for 1 hour and 0.5 g of AIBN was added. The mixturewas purged for ˜15 min until all of the AIBN dissolved. The solution washeated to 75° C. under nitrogen for 16 hours.

The white reaction mixture was filtered by vacuum filtration to yield awhite powder. The powder was washed with isopropanol and dried undervacuum to yield 22.20 g of a white powder (91.7% yield).

Additional MAPTAC/n-butylmethacrylate/methacrylamide terpolymers wereprepared by a similar method beginning with the following ratios ofstarting monomers: MAPTAC/n-butylmethacrylate/methacrylamide=20/20/60and 30/10/60.

Example 29 Synthesis of TMAEAC/n-Decylacrylamide/Acrylamid Terpolymer33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of n-decylacrylamide, and 8.06 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The reaction mixture was precipitated into 1200 mL of ethyl acetate. Thefine precipitate was filtered by vacuum filtration to yield a stickyyellow material. The light yellow solid was dissolved in methanol andprecipitated into 1200 mL of ethyl acetate. The precipitate was filteredby vacuum filtration to yield 10.85 g of a sticky, slightly yellowpowder (44.8% yield).

Example 30 Synthesis of TMAEAC/N-t-Butylacrylamide/Acrylamide Terpolymer33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of N-t-butylacrylamide, and 8.06 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of a clear colorless solution with asmall amount of white sticky solid. The clear solution was precipitatedinto 1200 mL of ethyl acetate. The white precipitate was filtered,dissolved in water, and lyophilized to yield 6.65 of a white powder(27.5% yield).

Example 31 Synthesis of TMAEAC/Styrene/Acrylamide Terpolymer 33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of styrene, and 8.06 g of acrylamide. The solution waspurged with nitrogen for 1 hour and 0.5 g AIBN was added. The mixturewas purged for ˜15 minutes until all of the AIBN dissolved. The solutionwas heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of a clear colorless solution and a whitesolid. The clear solution was disgarded. The solid was dissolved inmethanol, and precipitated into ethyl acetate (1200 mL). A whiteprecipitate formed which settled out of the solution as a sticky whitesolid. The ethyl acetate was decanted and the solid dried by passingnitrogen through the flask. The solid was dissolved in water andlyophilized to yield 18.14 g of a fine white powder (75% yield).

Example 32 Synthesis of TMAEAC/n-Butylacrylate/Acrylamide Terpolymer33133/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of n-butylacrylate, and 8.06 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The reaction mixture consisted of a clear colorless solution and a whitechunky solid. The solution phase was disgarded and the white soliddissolved in water, filtered and lyophilized to yield 12.84 of a finewhite powder (53.1% yield).

Example 33 Synthesis of TMAEAC/n-Decylacrylate/Acrylamide Terpolymer33/33133

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of n-decylacrylate, and 8.06 g of acrylamide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The white reaction mixture was precipitated into 1200 mL of ethylacetate. The turbid solution was decanted and the polymer was dried withnitrogen, dissolved in water, and lyophilized to yield 8.79 g of finewhite powder (36.3% yield).

Example 34 Synthesis of TMAEAC/t-Butylacrylate/Acrylamide Terpolymer33/33/33

To a 500 mL round-bottom, three-neck flask fitted with a thermocouple,reflux condenser, and septum was added 150 mL of isopropanol followed by16.13 g of a 50% aqueous solution of 2-trimethylammoniumethylacrylatechloride, 8.06 g of t-butylacrylate, and 8.06 g of acrylarnide. Thesolution was purged with nitrogen for 1 hour and 0.5 g AIBN was added.The mixture was purged for ˜15 minutes until all of the AIBN dissolved.The solution was heated to 75° C. under nitrogen for 16 hours.

The white reaction mixture was precipitated into 1200 mL of ethylacetate. The turbid solution was decanted and the polymer was dried withnitrogen, dissolved in water, and lyophilized to yield 6.51 g of finewhite powder (26.9% yield).

Example 35 In vitro activity of selected polymers against C. Parvuminfectivity

Confluent MDBK cell monolayers were grown on 16 well slides, andinfected with 5×10⁵ of C. parvum oocysts per well. Various dilutions ofthe test reagents in dimethylsulfoxide (DMSO) were added to themonolayers and cultures were incubated at 37° C.(8% CO₂) for 48 hours.The level of C parvum infections was determined and analysed by anindirect immunofluorescence (IF) assay at 48 hours. Anti-C. parvumsporozoite rabbit serum (1:1000) was used as the primary antibody, andfluorscein-conjugated anti rabbit goat serum (1:100) was used as thesecondary antibody. Each dilution was tested in quadruple, and eachassay was performed at least two times. The monolayers were methanolfixed and, after IF labelling, the number of parasites observed in 10high power fields (HPF) per well in each of the four wells per dilutionwas counted, statistically analysed and compared with infected wellswhich contained DMSO only. Paromomycin was used as the positive controldrug. The results are presented in the following Table.

TABLE Concentration Polymer (mg/mL) % Inhibitionpoly(TMAEMC-co-styrene)25/75, 0.1 91.7 Example 7 0.033 83.2 0.011 38.90.0037 3.95 poly(TMAEMC-co-N- 10 100 t-butylacrylamide), 15/85 Example13 1.0 100 0.1 59.1 0.01 38.0 poly(MAPTAC-co-N- 10 100n-decylacrylamide), 40/60 Example 14 1.0 100 0.1 64.3 0.01 35.5poly(MAPTAC-co-N- 10 70.2 t-butylacrylamide-co-HEMA) 33/33/33 Example 171.0 57.4 0.1 52.1 0.01 18.4 Poly(TMAEMC-co- 0.1 91.35heptafluorobutylacrylate60/40, Example 10 0.033 53.0 0.011 23.5 0.00374.2 paromomycin 2 79.4

EQUIVALENTS

Those skilled in the art will recognize or be able to ascertain using nomore than routine experimentation many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed in the scope of the following claims.

What is claimed is:
 1. A method for treating a microbial infection in amammal, comprising the step of administering to the mammal atherapeutically effective amount of a homopolymer characterized by apolymerized monomer represented by Formula I:

wherein R is a hydrogen atom or a methyl or ethyl group, X is a covalentbond or a CH₂ group, Y is an oxygen atom or an NH or CH₂ group, Z is analiphatic spacer group, and R₁, R₂ and R₃ are each, independently, ahydrogen atom, a normal or branched, substituted or unsubstitutedC₁-C₁₈-alkyl group, aryl or arylalkyl group.
 2. The method of claim 1wherein the mammal is a human.
 3. The method of claim 1 wherein Z is anormal or branched C₂-C₁₂-alkylene group or a C₂-C₁₂-alkylene groupinterrupted at one or more points by a heteroatom.
 4. A method fortreating a microbial infection in a mammal, comprising the step ofadministering to the mammal a therapeutically effective amount of acopolymer of a polymerized neutral hydrophobic monomer and a polymerizedmonomer represented by Formula I:

wherein R is a hydrogen atom or a methyl or ethyl group, X is a covalentbond or a CH₂ group, Y is an oxygen atom or an NH or CH₂ group, Z is analiphatic spacer group, and R₁, R₂ and R₃ are each, independently, ahydrogen atom, a normal or branched, substituted or unsubstitutedC₁-C₁₈-alkyl group, aryl or arylalkyl group.
 5. The method of claim 4wherein Z is a normal or branched C₂-C₁₂-alkylene group or aC₂-C₁₂-alkylene group interrupted at one or more points by a heteroatom.6. The method of claim 5 wherein the heteroatom is a nitrogen, oxygen orsulfur atom.
 7. The method of claim 4 wherein at least one of R₁, R₂ andR₃ is an aryl group, a benzyl group or a normal or branched, substitutedor unsubstituted C₁-C₁₈-alkyl group.
 8. The method of claim 4 whereinthe neutral hydrophobic monomer comprises a straight chain or branched,substituted or unsubstituted C₃-C₁₈-alkyl group, an aryl group or anaralkyl group.
 9. The method of claim 8 wherein the hydrophobic monomeris selected from the group consisting of styrene, N-isopropylacrylamide,N-t-butylacrylamide, N-n-butylacrylamide, heptafluorobutylacrylate,N-n-decylallylamine, N-n-decylacrylamide, pentafluorostyrene,n-butylacrylate, t-butylacrylate, n-decylacrylate,N-t-butylmethacrylamide, n-decylmethacrylate, n-butylmethacrylate,n-decylacrylate, and t-butylacrylate.
 10. A method for treating amicrobial infection in a mammal, comprising the step of administering atherapeutically effective amount of a terpolymer of a polymerizedmonomer having an amino group or an ammonium group attached to thepolymer backbone by an aliphatic spacer group, a polymerized neutralhydrophobic monomer and a polymerized neutral hydrophilic monomer. 11.The method of claim 10 wherein the terpolymer is administered orally andthe mammal is a human.
 12. The method of claim 11 wherein the monomerhaving an amino group or an ammonium group attached to the polymerbackbone by an aliphatic spacer group is represented by Formula I:

wherein R is a hydrogen atom or a methyl or ethyl group, X is a covalentbond, a carbonyl group or a CH₂ group, Y is an oxygen atom or an NH orCH₂ group, Z is an aliphatic spacer group, and R₁, R₂ and R₃ are each,independently, a hydrogen atom, a normal or branched, substituted orunsubstituted C₁-C₁₈-alkyl group, aryl or arylalkyl group.
 13. Themethod of claim 12 wherein X is a covalent bond or a CH₂ group.
 14. Themethod of claim 13 wherein the neutral hydrophilic monomer isacrylamide, methacrylamide, N-(2-hydroxyethyl)acrylamide and(2-hydroxyethyl)methacrylate.
 15. The method of claim 13 wherein Z is anormal or branched C₂-C₁₂-alkylene group or a C₂-C₁₂-alkylene groupinterrupted at one or more points by a heteroatom.
 16. A method fortreating a microbial infection in a mammal, comprising the step ofadministering to the mammal a therapeutically effective amount of apolymer comprising polymerized monomers, said monomers being representedby Formula I:

wherein R is a hydrogen atom or a methyl or ethyl group, X is a covalentbond or a CH₂ group, Y is an oxygen atom or an NH or CH₂ group, Z is analiphatic spacer group, and R₁, R₂ and R₃ are each, independently, ahydrogen atom, a normal or branched, substituted or unsubstitutedC₁-C₁₈-alkyl group, aryl or arylalkyl group.
 17. The method of claim 16wherein the mammal is a human.
 18. The method of claim 16 wherein Z is anormal or branched C₂-C₁₂-alkylene group or a C₂-C₁₂-alkylene groupinterrupted at one or more points by a heteroatom.
 19. A method fortreating a gastrointestinal infection in a human, comprising the step oforally administering to the human a therapeutically effective amount ofa polymer characterized by a polymerized monomer having an amino groupor an ammonium group attached to the polymer backbone by an aliphaticspacer group.
 20. A method for treating a gastrointestinal infection ina mammal, comprising the step of orally administering to the mammal atherapeutically effective amount of a polymer comprising polymerizedmonomers, said monomers being represented by Formula I:

wherein R is a hydrogen atom or a methyl or ethyl group, X is a covalentbond or a CH₂ group, Y is an oxygen atom or an NH or CH₂ group, Z is analiphatic spacer group, and R₁, R₂ and R₃ are each, independently, ahydrogen atom, a normal or branched, substituted or unsubstitutedC₁-C₁₈-alkyl group, aryl or arylalkyl group.